Cell & molecular analytical methods, visualizing cells. 1. Write the name of the

Question

Cell & molecular analytical methods, visualizing cells. 1. Write the name of the most appropriate technique that you can use to answer the question. Briefly explain (1-2 sentences) how this technique will answer the question. a. What DNA repair genes are expressed after exposing yeast to UV radiation? b. What DNA sequences does the engrailed transcription factor bind to in Drosophila? c. Does the amount of Hsp70 protein increase in E. coli following heat shock? You would like to examine the structure of the nuclear envelope, including the nuclear pores. What type of microscopy technique would you use and why?

Explanation / Answer

Ans1 a : Nucleotide-excision repair systems have been found in eukaryotes, particularly in yeasts. In yeasts, as in E. coli, several genes involved in DNA repair (called RAD genes for radiation sensitivity) have been identified by the isolation of mutants with increased sensitivity to UV light.

Ans 1.b An enzyme system that accurately initiates transcription of the engrailed gene has been prepared from Drosophila embryos. The system has been separated chromatographically into two fractions, both of which are required for specific engrailed transcription. DNase footprint and competition analysis detected at least two sequence-specific DNA-binding proteins in one of these two fractions. Together, these proteins bind to eight regions within 400 bp of the transcription initiation sites. Most of the regions containing these binding sites are required for manimal engrailed transcription in vitro. In addition, a region downstream from the initiation sites and within the first 40 residues of the transcription unit is essential for transcription.

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